PROJECT DESCRIPTION ? JSK Therapeutics (JSKT) Direct to Phase II Application Nitric oxide (NO) is a potentially powerful therapy against cancer. Spontaneous NO-generating drugs cannot be used to treat malignancies because of the pleiotropic effects of NO. In particular, NO is a potent vasodilator with resultant hypotension. In order to make NO-based cancer therapy possible, it is necessary to develop agents that deliver NO preferentially to malignant cells. In collaboration with Dr. Larry Keefer at the National Cancer Institute, Dr. Paul Shami has developed such a drug using compounds that release NO upon interaction with glutathione (GSH) in a reaction catalyzed by Glutathione S-Transferases (GST). This drug design exploits the over-expression of GST in malignant cells. A lead compound was identified as the most active of this family: O2-(2,4-dinitrophenyl) 1-[(4-ethoxycarbonyl) piperazin-1-yl]diazen-1-ium-1,2-diolate, or JS- K. JS-K has potent and broad-spectrum anti-cancer activity in vitro and in vivo. Because of challenging solubility and stability properties, the Shami lab has developed a formulation for JS-K using Pluronic P123 micelles. JS-K formulated in P123 micelles (P123/JS-K) was well tolerated in a non-GLP dog toxicology study without the induction of hypotension. JS-K is directly cytotoxic to malignant cells and also an inhibitor of angiogenesis. JS-K is a very efficient consumer of the anti-oxidant tripeptide glutathione (GSH), rendering malignant cells particularly susceptible to oxidative stress. JS-K is not toxic towards normal hematopoietic cells. JS-K is synergistic with cytarabine against acute myeloid leukemia (AML) and with bortezomib against multiple myeloma (MM). Recent data suggest that JS-K may sensitize MM cells to the cytotoxic effect of natural killer (NK) cells, raising the possibility of synergistic combinations with novel immunomodulatory agents. One kilogram of JS-K Active Pharmaceutical Ingredient was produced under cGMP conditions. JSK Therapeutics (JSKT) was founded to lead commercialization of JS-K. The United States Food and Drug Administration has granted orphan drug designation for JS-K for the indications of AML and MM. JSKT plans to develop JS-K initially for the treatment of AML and MM. The objective of this project is to complete pre-clinical animal toxicology studies needed to file an Investigational New Drug (IND) application with the FDA. In 4 aims, an analytical method for the measurement of JS-K in biologic matrices will be developed and validated; GLP toxicology studies in rats and dogs using a schedule of administration that reproduces the prospective clinical schedule will be conducted; GLP lung and central nervous system toxicology studies will be conducted in rats; and a GLP cardiovascular system toxicology study will be conducted in dogs. At the completion of this project, JSKT will submit an IND application to the FDA in order to start Phase I clinical trials for relapsed/refractory AML and MM. This will allow partnering with an established pharmaceutical company or obtaining venture investment in order to complete the clinical development and commercialization of JS-K, thus adding a potent new drug to our armamentarium aimed at diminishing the pain and suffering associated with cancer.